http://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
http://www.ahs.ac.cn/images/0513-353X/images/top-banner2.jpg|#|甘蓝
http://www.ahs.ac.cn/images/0513-353X/images/top-banner3.jpg|#|菊花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner4.jpg|#|灵芝
http://www.ahs.ac.cn/images/0513-353X/images/top-banner5.jpg|#|桃
http://www.ahs.ac.cn/images/0513-353X/images/top-banner6.jpg|#|黄瓜
http://www.ahs.ac.cn/images/0513-353X/images/top-banner7.jpg|#|蝴蝶兰
http://www.ahs.ac.cn/images/0513-353X/images/top-banner8.jpg|#|樱桃
http://www.ahs.ac.cn/images/0513-353X/images/top-banner9.jpg|#|观赏荷花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner10.jpg|#|菊花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner11.jpg|#|月季
http://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2020, Vol. 47 ›› Issue (3): 432-444.doi: 10.16420/j.issn.0513-353x.2019-403

• 研究论文 • 上一篇    下一篇

柑橘抗溃疡病转录因子CsAP2-09互作蛋白筛选与分析

祁静静*,窦万福*,张庆雯,胡安华,陈善春,雷天刚,彭爱红,许兰珍,姚利晓,何永睿**,李 强**   

  1. 西南大学/中国农业科学院柑桔研究所,国家柑桔工程技术研究中心,重庆 400712
  • 出版日期:2020-03-25 发布日期:2020-03-25
  • 基金资助:
    国家重点研发计划项目(2018YFD1000300);中央高校基本科研业务费项目(SWU115025);广西科技重大专项(桂科AA18118046-6);国家现代农业产业技术体系建设专项资金项目(CARS-26);重庆市社会事业与民生体系保障科技创新专项(cstc2016shms-ztzx80001,cstc2017shms-xdny80051)

Interacting Protein Screening and Analysis of CsAP2-09 — A Citrus Bacterial Canker Related Transcription Factor

QI Jingjing*,DOU Wanfu*,ZHANG Qingwen,HU Anhua,CHEN Shanchun,LEI Tiangang,PENG Aihong,XU Lanzhen,YAO Lixiao,HE Yongrui**,and LI Qiang**   

  1. Citrus Research Institute,Southwest University/Chinese Academy of Agricultural Sciences,National Citrus Engineering Technology Research Center,Chongqing 400712,China
  • Online:2020-03-25 Published:2020-03-25

摘要: 以前期获得的柑橘抗溃疡病的正调控转录因子基因 CsAP2-09 超表达植株为材料,利用 GST融合蛋白沉降技术(GST pull-down)联合液相色谱串联质谱(LC–MS/MS)方法筛选并鉴定 CsAP2-09的互作蛋白。首先原核表达并纯化 GST-CsAP2-09 作为诱饵蛋白,然后与 CsAP2-09 超表达植株叶片总蛋白孵育、洗脱后进行 SDS-PAGE 验证,最后进行 LC–MS/MS 鉴定。得到 17 个互作蛋白,将蛋白进行注释、GO 分析、KEGG 分析,发现其中 5 个可能与植物抗病相关(如过氧化氢酶 Cs3g27280)。构建这 17个蛋白的互作网络,其中 14 个蛋白的互作关系得到已有数据的支持。对 CsAP2-09 与互作蛋白的共表达分析发现 CsAP2-09 超表达引起了 16 个蛋白表达上调和 1 个蛋白表达下调。

关键词: 柑橘, 柑橘溃疡病, GST pull-down, CsAP2-09, AP2/ERF, 互作蛋白

Abstract: CsAP2-09 is a positive transcription factor involved in the citrus bacterial canker resistance. In this study,the GST pull-down and LC–MS/MS strategies were used for the screening and network analysis of the interacting proteins of CsAP2-09. Firstly,the fusion proteins GST-CsAP2-09 were expressed and purified for the next GST pull-down. Then GST-CsAP2-09 was fixed on the GST-beads and incubated with total proteins extracted from CsAP2-09 over-expression plant. The interacting proteins of CsAP2-09 were detected by LC–MS/MS. From the annotation,GO and KEGG,17 interacting proteins were obtained,5 of which were reported to be involved in bacterial resistance,e.g. Cs3g27280(CAT)-a catalase.The interacting network was constructed,among which 14 of 17 interacting proteins were involved. Sixteen proteins were positively regulated by CsAP2-09 in the transgenic plant while 1 protein was negatively regulated.

Key words: citrus, citrus bacterial canker(CBC), GST pull-down, CsAP2-09, AP2/ERF, interacting protein

中图分类号: